Pathology Residency Training Program
HLA Rotation – Objectives and Goals
Measurement of HLA has provided the basis for allowing successful bone marrow
transplantation and solid organ transplantation. Identification of a patient’s HLA-type initiates
the process, and typing of potential donors commences the donor selection process. Crossmatching potential donor and recipient is utilized to confirm compatibility. Factors for
consideration include: antibodies in the recipient directed against HLA components, which
could result in hyper-acute rejection of the graft or long-term graft rejection.
The overall goal of the HLA Laboratory is to provide the analyses and interpretations to help
with proper donor selection; several specific studies are utilized to accomplish this goal.
The HLA Laboratory’s Role in Transplantation:
To accurately, and in a timely manner, determine:
1. HLA-types of the recipient and potential donors
a. Serologic Determination
b. Molecular Determination
2. Panel-reactive antibodies (PRA) (anti-HLA antibodies)
a. CDC Determination
b. Fluorescence Determination
3. Cross-match compatibility
It is anticipated that the Resident will become sufficiently familiar with the assays:
1. To understand proper test selection, for the type of transplant to be performed.
2. To provide a reasonable level of troubleshooting of failed or confusing studies.
3. To be able to provide appropriate interpretation of the test results, to allow for
appropriate donor selection.
HLA Laboratory Assays: ACH
The HLA laboratory housed at ACH primarily provides testing for solid organ transplantation
(primarily renal allografts).
Tests Performed Include:
1. Class I ABC type by serologic testing (complement-dependent-cytotoxicity [CDC]
technique)
2. Class II DR DQ type by serologic testing (CDC technique)
3. PRA by serologic testing (CDC technique and AHG-enhanced / DTT-treated [as needed])
4. Cross-matching by serologic CDC technique – T lymphocyte and B lymphocyte
specificities (cells separated using magnetic microspheres)
5. PRA analysis screen by fluorescence cytometry – Percent positive on T lymphocytes and
B lymphocytes (HLA bead technology – T lymphocyte and B lymphocyte specificities –
identified by CD3 and CD19 monoclonal antibodies, respectively)
6. PRA single antigen determination by fluorescence cytometry
7. PRA specific antibody detection by Luminex (HLA bead technology with individual specific
HLA antigens)
8. Cross-matching by fluorescence cytometry – T lymphocyte and B lymphocyte specificities
(identified by CD3 and CD19 monoclonal antibodies, respectively)
9. HLA-B27 identification (currently using HLA-ABC Class I trays, rather than B27-specific
trays)
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HLA Laboratory Assays: UAMS
The HLA laboratory housed at UAMS provides testing primarily for the adult bone marrow
transplant program.
Tests Performed Include:
1. Class I ABC type serologic testing (CDC technique)
2. Class I ABC high resolution SSP-PCR
3. Class II DR DQ high resolution SSP-PCR
4. PRA by serologic testing (CDC technique / heat-treated [as needed])
5. HLA-B27 identification (currently using HLA-ABC Class I trays, rather than B27-specific
trays)
Future Assays Include:
1. Kir gene analyses by PCR
2. Cylex analysis of immunosuppressive therapy
3. Minor Histocompatibility Assays
To accomplish the goals: The Training Program will be structured to maximize the Resident’s
exposure to specific technical procedures and appropriate selection thereof, troubleshooting of
assays, and interpretation of assays. This will be accomplished by 4-8 weeks of Resident
participation in the laboratory. The Resident will take a written “Pre-Test” to assess potential
weaknesses, which can be concentrated upon during the allotted timeframe. The Resident will
directly interact with the technicians, for learning the procedures. The Resident will be
expected to type an unknown specimen by serologic and SSP-PCR techniques (the Resident
can use his/her own blood), measure PRA by serologic and Fluorescence Cytometry (the
Resident can use his/her own blood), and perform cross-match with PRA positive and negative
sera (the Resident can use his/her own blood). The Resident will have specific didactic
sessions each week with Dr. Harville, to discuss the theory and practical applications of HLA
testing. Specific cases will be reviewed and discussed. The Resident will be expected to do
his/her own outside reading. At the end of the training period, the Resident will take a “PostTest” to assess what has been learned. The Resident will be expected to write short
descriptions of 1) a patient receiving a solid organ transplant and 2) a patient receiving a bone
marrow transplant. The Resident will discuss the tests used, the results of the tests, how
donor selection was influenced by the specific testing, and outcomes (if known). The written
assignment will require review of laboratory material, and the discussions should reflect the
Resident’s understanding of the material, e.g., why a test was selected, problems with the
testing, how the results were determined, and how donor selection was influenced. The
written assignment will be due at the end of the rotation.
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